衰老途径参与大鼠动脉导管闭合机制的初步研究  

Primary study on the mechanism of ductus arteriosus closure in rats:impact of aging pathway

作　　者：尹惠梅 杨栋[2] 张邢炜[2,3] 焦其彬[3] Yin Huimei;Yang Dong;Zhang Xingwei;Jiao Qibin(Department of Pediatrics,Affiliated Hospital of Hangzhou Normal University,Hangzhou 310000,China;Department of Cardiology,Affiliated Hospital of Hangzhou Normal University,Hangzhou 310000,China;Hangzhou Normal University,Hangzhou 311121,China)

机构地区：[1]杭州师范大学附属医院儿科,310000 [2]杭州师范大学附属医院心血管内科,310000 [3]杭州师范大学,311121

出　　处：《中华心血管病杂志》2020年第5期408-412,共5页Chinese Journal of Cardiology

基　　金：浙江省自然科学基金(LY15H020007);浙江省医药卫生科研基金(2017KY526);浙江省科学技术厅科技计划(2016C33188)。

摘　　要：目的初步探讨衰老途径在大鼠动脉导管闭合过程中的作用及其机制。方法Spraygue Dawley远交群大鼠30只,其中雌鼠20只,10~15周龄,体重270~330 g,采用随机数表进行随机雌雄配对后交配,受孕后备用。提取孕19 d(E19组)、21 d(E21组)胎鼠和新出生(Day0组)幼鼠的动脉导管原代平滑肌细胞(DASMC)进行培养,于培养48 h后采用实时荧光定量PCR(RT-PCR)检测各组细胞衰老相关标志物p16、21和53基因的转录水平。低氧培养新生大鼠DASMC 3 d后将细胞分为3组,即低氧对照组(G0组)、正常氧浓度处理3 h组(G3组)和正常氧浓度处理6 h组(G6组),干预后采用RT-PCR检测各组细胞p16、21和53基因的转录水平。提取新生大鼠DASMC分为3组,即低氧培养对照组、低氧+小干扰RNA(siRNA)培养组和正常氧浓度培养组,通过Transwell实验检测DASMC迁移能力。结果E19组胎鼠DASMC的p16、21和53基因的转录水平均高于Day0组新生幼鼠(P均<0.01),E21组胎鼠DASMC的p16、21和53基因转录水平亦均高于Day0组新生幼鼠(P均<0.01)。G0组新生大鼠DASMC的p16、21和53基因的转录水平均高于G3组(P<0.05或0.01),G0组新生大鼠DASMC的p16、21和53基因的转录水平亦均高于G6组(P均<0.01),G3组新生大鼠DASMC的p16、21和53基因的转录水平则均高于G6组(P均<0.05)。正常氧浓度培养组新生大鼠DASMC的迁移能力高于低氧培养对照组(P<0.01),低氧+siRNA培养组新生大鼠DASMC的迁移能力亦高于低氧培养对照组(P<0.01)。结论大鼠动脉导管闭合过程中DASMC的衰老标志物随大鼠出生而降低,而衰老途径可能通过抑制DASMC迁移影响动脉导管闭合。Objective To explore the role and mechanism of aging pathway in patent ductus arteriosus closure of rats.Methods Thirty outbreeding Sprague Dawley rats(20 females,10-15 weeks old,270-330 g)underwent random mating and conception.The primary Ductus Arteriosus smooth muscle cells(DASMCs)of pregnant 19 days(E19 group),21 days(E21 group)and newborn(Day0 group)fetus were extracted and cultured.mRNA expression of cell senescence related markers p16,21 and 53 genes in each group were detected by real-time fluorescent quantitative PCR(RT-PCR)after 48 hours culture.After hypoxic culture on DASMCs for 3 days,the DASMCs were divided into 3 groups:hypoxic control group(G0 group),3 hours normal oxygen concentration treatment group(G3 group)and 6 hours normal oxygen concentration treatment group(G6 group).After intervention,mRNA expression of p16,21 and 53 RT-PCR was detected.The DASMCs of newborn rats(Day0 group)were extracted and divided into 3 groups:low-oxygen culture control group,low-oxygen+siRNA culture group and normal oxygen concentration culture group.The DASMCs migration ability was tested experimentally by Transwell method.Result The mRNA levels of p16,21 and 53 in DASMCs were higher in E19 group than in Day0 group(all P<0.01),and the mRNA levels of p16,21 and 53 in DASMCs were also higher in E21 group than those in Day0 group(all P<0.01).The mRNA levels of p16,21 and 53 in DASMC were all higher in G0 group than those in G3 group(P<0.05 or 0.01),and the mRNA levels of p16,21 and 53 in DASMCs were all higher in G0 group than those in G6 group(all P<0.01),and the mRNA levels of p16,21 and 53 in DASMCs were all higher in G3 group than those in G6 group(all P<0.05).DASMCs migration ability of newborn rats was higher in normal oxygen concentration culture group than that in low-oxygen culture group(P<0.01),and DASMCs migration ability of newborn rats was also higher in low-oxygen+siRNA culture group than that in low-oxygen culture group(P<0.01).Conclusion The expression of senescence marker of DASMCs decreases with the b

关 键 词：动脉导管未闭 衰老 肌细胞 平滑肌 

分 类 号：R[医药卫生]